隐丹参酮调节JAK2/STAT3通路抑制胃癌耐药细胞增殖  

Cryptotanshinone Inhibits Proliferation of Drug Resistant Gastric Cancer Cells by Regulating JAK2/STAT3 Signaling Pathway

作　　者：曹叶芝 张琦[1,2] 汪玲虎 程玲 储俊[4] 于庆生[1,2] 吴文楷[1] 刘海韦 舒亚倩 CAO Yezhi;ZHANG Qi;WANG Linghu;CHENG Ling;Chu Jun;YU Qingsheng;WU Wenkai;LIU Haiwei;SHU Yaqian(Department of General Surgery,The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei Anhui China 230031;Institute of Traditional Chinese Surgery,Anhui Academy of Chinese Medicine,Hefei Anhui China 230031;Xin′an Medical Laboratory of Ministry of Education,Anhui University of Chinese Medicine,Hefei Anhui China 230031;Department of Internal Medicine Intensive Care Unit,The First Affiliated Hospital to Anhui University of Chinese Medicine,Hefei Anhui China 230031)

机构地区：[1]安徽中医药大学第一附属医院普外一科,安徽合肥230031 [2]安徽省中医药科学院中医外科研究所,安徽合肥230031 [3]安徽中医药大学第一附属医院内科重症监护室,安徽合肥230031 [4]安徽中医药大学新安医学教育部重点实验室,安徽合肥230031

出　　处：《中医学报》2024年第5期1036-1044,共9页Acta Chinese Medicine

基　　金：安徽省自然科学基金面上项目(2008085MH266)。

摘　　要：目的:基于Janus蛋白酪氨酸激酶2(Janus kinase 2,JAK2)/信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)信号通路探讨隐丹参酮(cryptotanshinone,CPT)对胃癌5-氟尿嘧啶耐药细胞(SGC-7901/5-FU细胞)的影响及作用机制。方法:取对数生长期的SGC-7901/5-FU细胞,分为对照组(只含细胞)、5-FU组(细胞+25μmol·L^(-1)的5-FU)、CPT组(细胞+60μmol·L^(-1)的CPT)、5-FU+CPT组(细胞+25μmol·L^(-1)的5-FU和60μmol·L^(-1)的CPT)。采用CCK-8法检测细胞增殖情况;免疫荧光染色检测细胞核形态;流式细胞术检测细胞凋亡率;qPCR检测细胞B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2关联X蛋白(Bcl-2 associated X protein,Bax)、生存素(Survivin)、JAK2、STAT3、多重耐药蛋白1(multidrug resistance protein 1,MDR1)基因表达水平;Western blot检测Bcl-2、Bax、Survivin、JAK2、p-JAK2、STAT3、p-STAT3、P-糖蛋白(P-glycoprotein,P-gp)表达水平。结果:CCK-8结果显示,5-FU或CPT干预24 h对细胞的抑制作用最显著;随着药物浓度增加,细胞抑制率逐渐升高。5-FU和CPT对SGC-7901/5-FU细胞的半抑制浓度(half maximal inhibitory concentration,IC 50)分别为26.94μmol·L^(-1)、63.45μmol·L^(-1)。免疫荧光结果显示,对照组细胞核结构完整,呈卵圆形;5-FU组、CPT组和5-FU+CPT组细胞核出现变形、皱缩和密集染色现象。与对照组比较,5-FU组、CPT组和5-FU+CPT组细胞凋亡率增加(P<0.05);与5-FU组比较,5-FU+CPT组细胞凋亡率增加(P<0.05)。与对照组比较,5-FU组、CPT组和5-FU+CPT组细胞Bcl-2 mRNA、Survivin mRNA水平降低(P<0.05);与对照组比较,CPT组、5-FU+CPT组细胞Bax mRNA水平升高(P<0.05),MDR1 mRNA水平降低(P<0.01);与5-FU组比较,5-FU+CPT组细胞Bcl-2 mRNA、MDR1 mRNA水平降低(P<0.05),Bax mRNA水平升高(P<0.01)。与对照组比较,CPT组、5-FU+CPT组细胞Bcl-2、Survivin、P-gp蛋白表达水平降低(P<0.05),Bax蛋白表达水平升高(P<0.05);与5-FU组比较,5-FU+CPT组细胞Bcl-2�Objective:To investigate the effect and mechanism of cryptotanshinone(CPT)on 5-fluorouracil-resistant gastric cancer cell(SGC-7901/5-FU cell)based on Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathways.Methods:SGC-7901/5-FU cells in logarithmic growth phase were taken.The cells were divided into control group(cells only),5-FU group(cells+25μmol·L^(-1)5-FU),CPT group(cells+60μmol·L^(-1) CPT),5-FU+CPT group(cells+25μmol·L^(-1)5-FU and 60μmol·L^(-1) CPT).Cell proliferation was detected by CCK-8 assay.The nuclear morphology was detected by immunofluorescence staining.The apoptosis rate was detected by flow cytometry.qPCR was used to detect B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax),Survivin,JAK2,STAT3 and multidrug resistance protein 1(MDR1)gene expression levels;The expression levels of Bcl-2,Bax,Survivin,JAK2,p-JAK2,STAT3,p-STAT3 and P-glycoprotein(P-gp)were detected by Western blot.Results:CCK-8 results showed that 5-FU or CPT treatment for 24 h had the most significant inhibitory effect on the cells.With the increase of drug concentration,the cell inhibition rate gradually increased.The half maximal inhibitory concentrations(IC 50)of 5-FU and CPT on SGC-7901/5-FU cells were 26.94μmol·L^(-1) and 63.45μmol·L^(-1) respectively.Immunofluorescence results showed that the nucleus structure of the control group was intact and oval.Nuclear deformation,shrinkage and dense staining were observed in 5-FU group,CPT group and 5-FU+CPT group.Compared with the control group,the apoptosis rates of 5-FU,CPT and 5-FU+CPT groups were increased significantly(P<0.05).Compared with 5-FU group,the apoptosis rate of 5-FU+CPT group was increased significantly(P<0.05).Compared with the control group,the expression levels of Bcl-2 mRNA and Survivin mRNA in 5-FU group,CPT group and 5-FU+CPT group were decreased significantly(P<0.05).Compared with the control group,the Bax mRNA level was increased significantly(P<0.05),and the MDR1 mRNA level was decreased significantl

关 键 词：隐丹参酮 胃癌 SGC-7901/5-FU细胞 JAK2/STAT3通路 细胞增殖 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]